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Paper-Chromatography.ppt
- 2. What is Chromatography? Chromatographyis a technique for separating mixtures into their components in order to analyze, identify, purify, and/or quantify the mixture or components. Separate • Analyze • Identify • Purify • Quantify Components Mixture
- 3. Uses for Chromatography Chromatographyis used by scientists to: • Analyze – examine a mixture, its components, and their relations to one another • Identify – determine the identity of a mixture or components based on known components • Purify – separate components in order to isolate one of interest for further study • Quantify – determine the amount of the a mixture and/or the components present in the sample
- 4. Uses for Chromatography Real-lifeexamples of uses for chromatography: • Pharmaceutical Company – determine amount of each chemical found in new product • Hospital – detect blood or alcohol levels in a patient’s blood stream • Law Enforcement – to compare a sample found at a crime scene to samples from suspects • Environmental Agency – determine the level of pollutants in the water supply • Manufacturing Plant – to purify a chemical needed to make a product
- 5. Definition of Chromatography DetailedDefinition: Chromatography is a laboratory technique that separates components within a mixture by using the differential affinities of the components for a mobile medium and for a stationary adsorbing medium through which they pass. Terminology: • Differential – showing a difference, distinctive • Affinity – natural attraction or force between things • Mobile Medium – gas or liquid that carries the components (mobile phase) • Stationary Medium – the part of the apparatus that does not move with the sample (stationary phase)
- 6. Simplified Definition: Chromatography separatesthe components of a mixture by their distinctive attraction to the mobile phase and the stationary phase. Explanation: • Compound is placed on stationary phase • Mobile phase passes through the stationary phase • Mobile phase solubilizes the components • Mobile phase carries the individual components a certain distance through the stationary phase, depending on their attraction to both of the phases Definition of Chromatography
- 7. Illustration of Chromatography Components Affinityto Stationary Phase Affinity to Mobile Phase Blue ---------------- Insoluble in Mobile Phase Black Red Yellow Mixture Components Separation Stationary Phase Mobile Phase
- 9. • Liquid Chromatography– separates liquid samples with a liquid solvent (mobile phase) and a column composed of solid beads (stationary phase) • Gas Chromatography – separates vaporized samples with a carrier gas (mobile phase) and a column composed of a liquid or of solid beads (stationary phase) • Paper Chromatography – separates dried liquid samples with a liquid solvent (mobile phase) and a paper strip (stationary phase) • Thin-Layer Chromatography – separates dried liquid samples with a liquid solvent (mobile phase) and a glass plate covered with a thin layer of alumina or silica gel (stationary phase) Types of Chromatography Types of Chromatography
- 10. (A) uses charge,(B) uses pores, and (C) uses covalent bonds to create the differential affinities among the mixture components for the stationary phase.
- 11. Principles of PaperChromatography • Capillary Action – the movement of liquid within the spaces of a porous material due to the forces of adhesion, cohesion, and surface tension. The liquid is able to move up the filter paper because its attraction to itself is stronger than the force of gravity. • Solubility – the degree to which a material (solute) dissolves into a solvent. Solutes dissolve into solvents that have similar properties. (Like dissolves like) This allows different solutes to be separated by different combinations of solvents. Separation of components depends on both their solubility in the mobile phase and their differential affinity to the mobile phase and the stationary phase.
- 12. Paper Chromatography Experiment WhatColor is that Sharpie?
- 13. Overview of theExperiment Purpose: To introduce students to the principles and terminology of chromatography and demonstrate separation of the dyes in Sharpie Pens with paper chromatography. Time Required: Prep. time: 10 minutes Experiment time: 45 minutes Costs: Less than $10
- 14. • 6 beakersor jars • 6 covers or lids • Distilled H2O • Isopropanol • Graduated cylinder • 6 strips of filter paper • Different colors of Sharpie pens • Pencil • Ruler • Scissors • Tape Materials List
- 15. Preparing the Isopropanol Solutions •Prepare 15 ml of the following isopropanol solutions in appropriately labeled beakers: - 0%, 5%, 10%, 20%, 50%, and 100%
- 16. Preparing the Chromatography Strips •Cut 6 strips of filter paper • Draw a line 1 cm above the bottom edge of the strip with the pencil • Label each strip with its corresponding solution • Place a spot from each pen on your starting line
- 17. Developing the Chromatograms •Place the strips in the beakers • Make sure the solution does not come above your start line • Keep the beakers covered • Let strips develop until the ascending solution front is about 2 cm from the top of the strip • Remove the strips and let them dry
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- 21. Observing the Chromatograms Concentrationof Isopropanol 0% 20% 50% 70% 100%
- 22. Black Dye Concentration ofIsopropanol 0% 20% 50% 70% 100% 1. Dyes separated – purple and black 2. Not soluble in low concentrations of isopropanol 3. Partially soluble in concentrations of isopropanol >20%
- 23. Blue Dye Concentration ofIsopropanol 0% 20% 50% 70% 100% 1. Dye separated – blue 2. Not very soluble in low concentrations of isopropanol 3. Completely soluble in high concentrations of isopropanol
- 24. Green Dye Concentration ofIsopropanol 0% 20% 50% 70% 100% 1. Dye separated – blue and yellow 2. Blue – Soluble in concentrations of isopropanol >20% 3. Yellow – Soluble in concentrations of isopropanol >0%
- 25. Red Dye 1. Dyesseparated – red and yellow 2. Yellow –soluble in low concentrations of isopropanol and less soluble in high concentrations of isopropanol Concentration of Isopropanol 0% 20% 50% 70% 100% 3. Red – slightly soluble in low concentrations of isopropanol, and more soluble in concentrations of isopropanol >20%
- 26. Alternative Experiments • Testdifferent samples: – Other markers, pens, highlighters – Flower pigments – Food Colors • Test different solvents: – Other alcohols: methanol, ethanol, propanol, butanol • Test different papers: – Coffee filters – Paper towels – Cardstock – Typing paper
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